mouse fibroblasts with Viromer® BLUE, Viromer® GREEN or Lipofectamine®. >> both Viromers ... primary mouse fibroblasts: transfection of siRNA. Lipocalyx ...
primary mouse fibroblasts: transfection of siRNA
VB-01LB-00, VB-01LB-01
VG-01LB-00, VG-01LB-01
Fig. 1: Comparative knockdown efficiency after siRNA transfection in primary mouse fibroblasts with Viromer® BLUE, Viromer® GREEN or Lipofectamine® >> both Viromers give a higher KD (>60%) compared to Lipofectamine (35%)
“And thank you very much for letting us test the Viromer system, we were satisfied with the efficiency of knockdown!”
Data from J. Holland, MDC Berlin (Germany)
Lipocalyx GmbH
Weinbergweg 23
06120 Halle
Germany
viromer-transfection.com viromer-transfection.com
primary human fibroblasts: transfection of siRNA
VB-01LB-00, VB-01LB-01
VG-01LB-00, VG-01LB-01
pictures taken at 2.5 x magnification
Fig. 2: Comparison test of transfection efficiency with a labeled siRNA (siGLO) in primary human fibroblasts using Viromer® BLUE, Viromer® GREEN and Lipofectamine® RNAiMAX
Lipocalyx GmbH
Weinbergweg 23
“I was pleasantly surprised about the transfection efficiency I saw with the Green product from my few trials. It was definitely much better than for RNAi max.”
Data from P. Holland, University of Oslo (Norway)
06120 Halle
Germany
viromer-transfection.com viromer-transfection.com
primary rat fibroblasts: transfection of siRNA
VB-01LB-00, VB-01LB-01
Material & methods “Transient gene silencing in UUOF (ndlr: primary cultures propagated from fibrotic kidney) was performed using pre-designed siRNA (13 nM) ON-TARGET plus SMARTpool reagents from Dharmacon (GE Life Sciences): Tgfb1 (L-091428-02) and a non-targeting control pool (D-001810-10). Fibroblasts were transfected with siRNA using Viromer blue (Lipocalyx) according to the manufacturer’s protocol in FBS-reduced media (1%) with antibiotics for 48 h.”
Fig. 3: from Fig. 8 in Smith et al. 2017 (b,c) The effect of Tgfb1 specific siRNAs on fibrogenic gene expression using qRT-PCR (b), αSMA levels and pSmad2/3 protein levels (c) in UUOF treated with FGF23 (1 ng/mL for 24 h), compared to a non-targeting control (NTC) pool. Representative Western blot of TGF-β1 silencing shown above.
Smith, ER et al. "FGF23 is synthesised locally by renal tubules and activates injury-primed fibroblasts." Scientific Reports 7 (2017). Lipocalyx GmbH
Weinbergweg 23
06120 Halle
Germany
viromer-transfection.com viromer-transfection.com
primary human fibroblasts: transfection of pDNA
VY-01LB-00, VY-01LB-01
Fig. 4: Overexpression of GFP in primary human fibroblasts after plasmid transfection with Viromer® YELLOW
“We are very satisfied with the performance of your product in our hands. Indeed, the difficult to transfect cell line of human primary fibroblasts that I was testing your Viromers on seem to be nicely responding to Viromer Yellow. I think your product will be highly appreciated around here. “
Data from E. Markkanen, University of Oxford (UK)
Lipocalyx GmbH
Weinbergweg 23
06120 Halle
Germany
viromer-transfection.com viromer-transfection.com
primary rat fibroblasts: transfection of pDNA Single and co-transfection of plasmid DNA in rat primary fibroblasts propagated from fibrotic kidney with Viromer® YELLOW (UUOF = unilateral ureteric obstruction fibroblasts)
VY-01LB-00, VY-01LB-01
Fig. 5: from Fig. 8 “Effects of FGF23 on UUOF are mediated by FGFR and activation of TGF-β1 signaling” in Smith et al. 2017 (d,e) The effect of pre-treatment with naringenin (Smad 3 inhibitor) on fibrogenic gene expression using qRT-PCR (d), αSMA levels and pSmad2/3 protein levels (e) in UUOF treated with FGF23 (1 ng/mL for 24 h), compared to a NTC pool. (f,g) The effect of transient overexpression of Smad7 (endogenous Smad inhibitor) on fibrogenic gene expression using qRT-PCR (f), and quantitative flow cytometric analysis of αSMA levels and pSmad2/3 (g) in UUOF treated with FGF23 (1 ng/mL for 24 h), compared to cells transfected with empty vector. Representative Western blot (upper left ‘f’) of Smad7-transfected UUOF. Suppression of Smad signalling was confirmed using a reporter assay as previously described (upper right ‘f’).
Smith, ER et al. "FGF23 is synthesised locally by renal tubules and activates injuryprimed fibroblasts." Scientific Reports 7 (2017).
Lipocalyx GmbH
Weinbergweg 23
06120 Halle
Germany
viromer-transfection.com viromer-transfection.com
Primary human fibroblasts: transfection of shRNA
VR-01LB-00, VR-01LB-01
Fig. 6 (from Fig.3 B in Simkova et al. 2016): Western blot analysis shows downregulation of asporin by shRNA in primary fibroblasts isolated from patients with breast cancer (CAF: cancer associated fibroblasts, cells alone) Use of Viromer® RED for transient asporin KD, change of medium after 12h
Simkova, D et al. "The dual role of asporin in breast cancer progression." Oncotarget 7.32 (2016): 52045.
Lipocalyx GmbH
Weinbergweg 23
06120 Halle
Germany
viromer-transfection.com viromer-transfection.com
